Genetic architecture of glucosinolate variation in Brassica napus

Kittipol, Varanya; He, Zhesi; Wang, Lihong; Doheny-Adams, Tim; Langer, Swen; Bancroft, Ian

doi:10.1016/j.jplph.2019.06.001

Cited by 38 publications

(45 citation statements)

References 49 publications

(62 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Previously, AtMAM1 was shown to be responsible for HPhe production in A. thaliana, and heterologous expression of AtMAM1 in E. coli enables HPhe production (Petersen et al, 2019b). Likewise, a MAM1 homolog in Brassica napus has been genetically linked to 2PE formation (Kittipol et al, 2019). Our data suggests that BvMAM1 has substrate specificity towards α-keto acids from both phenylalanine and methionine.…”

Section: Discussionmentioning

confidence: 53%

Engineering and optimization of the 2-phenylethylglucosinolate production inNicotiana benthamianaby combining biosynthetic genes fromBarbarea vulgarisandArabidopsis thaliana

Wang

Crocoll

Agerbirk

et al. 2020

Preprint

View full text Add to dashboard Cite

Among the glucosinolate (GLS) defense compounds characteristic of the Brassicales order, several have been shown to promote human health. This includes 2phenylethylglucosinolate (2PE) derived from homophenylalanine (HPhe). In this study, we used transient expression in Nicotiana benthamiana to validate and characterize previously predicted key genes in the 2PE biosynthetic pathway from Barbarea vulgaris and 2 demonstrate the feasibility of engineering 2PE production. We used genes from B. vulgaris and Arabidopsis thaliana, in which the biosynthesis of GLSs is predominantly derived from HPhe and dihomomethionine, respectively. The resulting GLS profiles partially mirrored GLS profiles in the gene donor plant, but in both cases the profiles in N. benthamiana were wider than in the native plants. We found that BvBCAT4 is a more efficient entry enzyme for biosynthesis of both HPhe and dihomomethionine and that MAM1 enzymes determine the chain-elongated profile. Co-expression of the chain elongation pathway and CYP79F6 from B. vulgaris with the remaining aliphatic GLS core pathway genes from A. thaliana, demonstrated the feasibility of engineering production of 2PE in N. benthamiana. Noticeably, the HPhe-converting enzyme BvCYP79F6 in the core GLS pathway belongs to the CYP79F subfamily, a family believed to have substrate specificity towards chain-elongated methionine derivatives. Replacing the B. vulgaris chain elongation pathway with a chimeric pathway consisting of BvBCAT4, BvMAM1, AtIPMI and AtIPMDH1 resulted in an additional 2-fold increase in 2PE production, demonstrating that chimeric pathway with genes from different species can increase flux and boost production in an engineered pathway. Our study provides a novel approach to produce the important HPhe and 2PE in a heterologous host. Chimeric engineering of a complex biosynthetic pathway enabled detailed understanding of catalytic properties of individual enzymes -a prerequisite for understanding biochemical evolution -and with biotechnological and plant breeding potentials of new-tonature gene combinations.

show abstract

Section: Discussionmentioning

confidence: 53%

Engineering and optimization of the 2-phenylethylglucosinolate production inNicotiana benthamianaby combining biosynthetic genes fromBarbarea vulgarisandArabidopsis thaliana

Wang

Crocoll

Agerbirk

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…However, the prominent role of the C2 copy of MYB28 is supported by a recent publication in which the A9 and C2 copies of MYB28 in Brassica napus are found to be responsible for glucosinolate accumulation, especially glucoraphanin, in leaves (Kittipol et al, 2019). Further analysis may include protein binding assays and analysis of progeny of back-crossed individuals with single mutations.…”

Section: Mutations In Myb28 Dna Binding Domainmentioning

confidence: 96%

“…This is further complicated by two additional copies of the close homologue, MYB29, on chromosome 3 (Bo3g004500) and chromosome 9 (Bo9g175680), which is known to work alongside MYB28 in regulating aliphatic glucosinolate biosynthesis (Gigolashvili et al, 2007;Hirai et al, 2007;Sonderby et al, 2007). The expression of the C2 copy is thought to be of most importance in regulating aliphatic glucosinolate biosynthesis in aerial organs (Kittipol et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

CRISPR-Cas9-mediated editing ofmyb28impairs glucoraphanin accumulation ofBrassica oleraceain the field

Neequaye

Lawrenson

et al. 2020

Preprint

View full text Add to dashboard Cite

We sought to quantify the role of MYB28 in the regulation of aliphatic glucosinolate biosynthesis and associated sulphur metabolism in field-grown B. oleracea with the use of CRISPR-Cas9-mediated gene editing technology. We describe the first characterised myb28 knockout mutant in B. oleracea, and the first DEFRA-regulated and approved CRISPR field trial in the UK to require compliance with the 2001/18 EU GMO directive. We report that knocking-out myb28 results in downregulation of aliphatic glucosinolate biosynthesis genes and reduction in accumulation of the methionine-derived glucosinolate, glucoraphanin, in leaves and florets of field-grown myb28 mutant broccoli plants. There were no significant changes to the accumulation of sulphate, S-methyl cysteine sulfoxide and indole glucosinolate in leaf and floret tissues.

show abstract

“…While GSL inclusion in diet affects health of livestock e.g., damaging liver, kidney, and thyroid gland and impairing fertility [ 96 ], this secondary metabolite could be useful as a cancer prevention and plant protection agent [ 97 , 98 ]. The importance of GSLs in plant defense has been demonstrated [ 99 ] and explains the increase in susceptibility to pests and diseases of the “00” (low seed erucic acid and GSL) rapeseed cultivars [ 100 ].…”

Section: Breeding For Economically Important Agronomic Traits Of mentioning

confidence: 99%

“…Over the at least 15 years, knowledge on metabolism of GSLs in Brassica crops and Arabidopsis, and the underlying genetic architecture has been achieved [ 97 , 98 , 100 , 101 ]. To date, there are at least 130 different classes of plant GSL, almost all from the Brassicales order, summarized by Blažević et al [ 102 ] and approximately 40 different GSLs synthesized in Arabidopsis [ 103 ].…”

Section: Breeding For Economically Important Agronomic Traits Of mentioning

confidence: 99%

The Use of Genetic and Gene Technologies in Shaping Modern Rapeseed Cultivars (Brassica napus L.)

Ton

Neik

Batley

2020

Genes

View full text Add to dashboard Cite

Since their domestication, Brassica oilseed species have undergone progressive transformation allied with the development of breeding and molecular technologies. The canola (Brassica napus) crop has rapidly expanded globally in the last 30 years with intensive innovations in canola varieties, providing for a wider range of markets apart from the food industry. The breeding efforts of B. napus, the main source of canola oil and canola meal, have been mainly focused on improving seed yield, oil quality, and meal quality along with disease resistance, abiotic stress tolerance, and herbicide resistance. The revolution in genetics and gene technologies, including genetic mapping, molecular markers, genomic tools, and gene technology, especially gene editing tools, has allowed an understanding of the complex genetic makeup and gene functions in the major bioprocesses of the Brassicales, especially Brassica oil crops. Here, we provide an overview on the contributions of these technologies in improving the major traits of B. napus and discuss their potential use to accomplish new improvement targets.

show abstract

Genetic architecture of glucosinolate variation in Brassica napus

Cited by 38 publications

References 49 publications

Engineering and optimization of the 2-phenylethylglucosinolate production inNicotiana benthamianaby combining biosynthetic genes fromBarbarea vulgarisandArabidopsis thaliana

Engineering and optimization of the 2-phenylethylglucosinolate production inNicotiana benthamianaby combining biosynthetic genes fromBarbarea vulgarisandArabidopsis thaliana

CRISPR-Cas9-mediated editing ofmyb28impairs glucoraphanin accumulation ofBrassica oleraceain the field

The Use of Genetic and Gene Technologies in Shaping Modern Rapeseed Cultivars (Brassica napus L.)

Contact Info

Product

Resources

About